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Publication : Single-copy transgenic mice with chosen-site integration.

First Author  Bronson SK Year  1996
Journal  Proc Natl Acad Sci U S A Volume  93
Issue  17 Pages  9067-72
PubMed ID  8799155 Mgi Jnum  J:34887
Mgi Id  MGI:82342 Doi  10.1073/pnas.93.17.9067
Citation  Bronson SK, et al. (1996) Single-copy transgenic mice with chosen-site integration [see comments]. Proc Natl Acad Sci U S A 93(17):9067-72
abstractText  We describe a general way of introducing transgenes into the mouse germ line for comparing different sequences without the complications of variation in copy number and insertion site. The method uses homologous recombination in embryonic stem (ES) cells to generate mice having a single copy of a transgene integrated into a chosen location in the genome. To test the method, a single copy murine bcl-2 cDNA driven by either a chicken beta-actin promoter or a human beta-actin promoter has been inserted immediately 5' to the X-linked hypoxanthine phosphoribosyltransferase locus by a directly selectable homologous recombination event. The level of expression of the targeted bcl-2 transgene in ES cells is identical in independently isolated homologous recombinants having the same promoter yet varies between the different promoters. In contrast, the expression of bcl-2 transgenes having the same (chicken beta-actin) promoter varies drastically when they are independently integrated at random insertion sites. Both promoters direct broad expression of the single-copy transgene in mice derived from the respective targeted ES cells. In vitro and in vivo, the human beta-actin promoter consistently directed a higher level of transgene expression than the chicken beta-actin promoter.
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